Biochemical and structural characterization of CYP109A2, a vitamin D₃ 25‐hydroxylase from Bacillus megaterium

Abstract

Cytochrome P450 monooxygenase CYP109A2, EC 1.14.14.1, UniProt ID: D5DF88, Ferredoxin, UniProt ID: D5DFQ0, cytochrome P450 reductase, EC 1.8.1.2, UniProt ID: D5DGX1.

Plant growth-promoting activities and genomic analysis of the stress-resistant Bacillus megaterium STB1, a bacterium of agricultural and biotechnological interest

“…In addition, the genome of strain STB1 contains several cytochrome p450 encoding genes ( Table S7 ), including a cytochrome P450/NADPH--P450 reductase homolog (C0569_02245) (98.1 % identity to B. megaterium ATCC 14581 CYP102A1) involved in fatty acid hydroxylation [43]; a CYP109E1 homolog (C0569_04895) (98.5 % identity to B. megaterium DSM 319 CYP109E1) involved in the hydroxylation of steroids, vitamin D3 (cholecalciferol), cholesterol, statins and terpenes [[44], [45], [46],47]; a CYP106A1 homolog (C0569_20645) (95.4 % identity to B. megaterium DSM 319 CYP106A1) involved in the hydroxylation of the pentacyclic triterpene 11-keto-β-boswellic acid and several steroids [48,49]; a CYP109A2 homolog (C0569_21565) (97.3 % identity to B. megaterium DSM 319 CYP109A2) involved in vitamin D3 hydroxylation [50]; and a unknown cytochrome P450 (C0569_21365) with 96 % identity to a B. megaterium DSM 319 WP_080514752.1 cytochrome P450.…”

Characterization and structure‐guided engineering of the novel versatile terpene monooxygenase CYP109Q5 from Chondromyces apiculatus DSM436

“…Crystals of CYP109Q5 were in space group P 2 1 1. The structure was solved using MOLREP (Vagin and Teplyakov, ), using CYP109A2 (5OFQ, Abdulmughni et al ., ) from Bacillus megaterium as the search model. The solution contained one monomer in the asymmetric unit and the solvent content was 42.4%.…”

The “beauty in the beast”—the multiple uses of Priestia megaterium in biotechnology

“…Furthermore, enzymes of vitamin B 12 and heme biosynthesis (Biedendieck et al 2010;Leech et al 2003;Mobius et al 2010;Moore et al 2013a;Moore et al 2014), reductive dehalogenases (Payne et al 2015), and the model green fluorescent protein (GFP) (Biedendieck et al 2007b;Biedendieck et al 2007c;Gamer et al 2009;Stammen et al 2010a;Stammen et al 2010b) complete the picture. Finally, the challenging cytochrome P450 enzymes, catalyzing for example stereospecific hydroxylation of steroids or vitamin D 3 , are naturally encoded by different P. megaterium genomes and were recombinantly produced using this bacterium (Abdulmughni et al 2017a;Abdulmughni et al 2017b;Bleif et al 2012;Brill et al 2014;Ehrhardt et al 2016;Gerber et al 2015).…”